Ionic aqueous polysaccharide compositions

ABSTRACT

Disclosed herein are ionic aqueous compositions useful as nasal passage washes to aid in the resorption of edema of the respiratory mucosa, e.g., the nasal mucosa, for the treatment of respiratory tract and/or respiratory mucosal-related conditions, including, e.g., chronic rhinosinusitis, sinusitis, allergic rhinitis and nasal polyps. The composition includes an ionic aqueous solution and algae-derived constituents, such as branched, sulfated polysaccharides having an average molecular weight greater than 4 kDa and comprising L-fucose and sulfate ester groups or extracts from brown algae. Also disclosed are methods and pharmaceutical compositions for treating respiratory tract and/or respiratory mucosal-related conditions, including, e.g., chronic rhinosinusitis.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. application Ser. No. 15/100,187filed 28 Nov. 2014; which is a US National Phase Application of PCTInternational Application No. PCT/EP14/75983 filed 28 Nov. 2014; whichclaims the benefit of U.S. Provisional Application Ser. No. 61/911,071filed 3 Dec. 2013; each of which is incorporated herein by reference inits entirety.

BACKGROUND OF THE INVENTION

Sinonasal complaints affect at least 15% of the population in the UnitedStates of America with predictable patient morbidity and financialimpact. Importantly, any modality that can alleviate sinonasalcomplaints warrants serious attention. Among various treatments utilizedin this setting, nasal irrigation is considered a simple, inexpensiveand beneficial approach that can be applied in concert with existingprescribed medications such as steroids, anti-histamines,vasoconstrictors and other drugs or as part of treatment modalities thathelp alleviate sinonasal symptoms such as nasal obstruction, rhinorrheaand others.

The principal functions of the nasal passages are the regulation of theinspiratory air flow, the heating and humidification of inspired air andthe elimination of foreign elements (dust, microbial germs). The ciliateepithelium, which covers the nasal mucosa, plays a vital role in itsdefense from infections. The vibratile cilia of this epithelium areimmersed in protective mucus, which surrounds them, and their purpose isto push this mucous, together with any external impurities, posteriorly,i.e., towards the pharynx, by their motions.

Seawater-based compositions currently available for rinsing the nasalpassages are generally prepared so that they are isotonic with the nasalmucosa. These compositions usually consist of a mixture of seawater anddeionized water, such that the per volume fraction of seawater in thismixture is approximately one-third. Alternatively, the compositionsconsist of increased per volume seawater fractions resulting inhypertonic solutions. These are believed to have better cleansingproperties due to the generation of osmotic gradients that allow formore efficacious mucus removal in comparison to isotonic solutions.

However, and despite their widespread use for cleansing and symptomaticrelief of sinonasal conditions, these solutions, typically packaged inthe form of aerosol sprays, provide little or no therapeutic benefitsagainst the causes of the disease. Furthermore, and in cases where theseawater compositions are complemented with additional ingredientsthought to improve their action, such as exogenous salts, anti-septicanti-bacterial agents, amino acids, essential oils and other agents,they suffer from limited industrial applicability due to variousfactors. These include poor water solubility of added ingredients, labelrestrictions and/or burning sensations (e.g. in the case of essentialoils; these cannot be used in children according to WHO guidelines),limited compatibility of added ingredients to the high salt contentpresent in (hypertonic) solutions, increased cost or other limitations.

For this reason, better seawater and/or saline solutions with improvedefficacy and negligible side effects are sought.

SUMMARY OF THE INVENTION

As described below, the present invention features compositions fornasal administration and methods of treating respiratory tract and/orrespiratory mucosal-related conditions.

In one aspect, a composition includes an ionic aqueous solution and asulfated polysaccharide having an average molecular weight greater than4 kDa and comprising L-fucose and sulfate ester groups. In oneembodiment, the composition is formulated for nasal administration.

In one embodiment, the sulfated polysaccharide is derived from anextract isolated from brown algae. In another embodiment, the brownalgae is Undaria pinnafitida. In another embodiment, the sulfatedpolysaccharide is fucoidan. In yet another embodiment, the sulfatedpolysaccharide is in a range of about 0.1 to about 10% weight content ofthe composition.

In another embodiment, the composition further includes an extract of ablue/green algae, e.g., such as an extract isolated from Spirulinaplatensis. In one embodiment, the blue/green algae extract is in a rangeof about 0.1 to about 10% weight content of the composition.

In yet another embodiment, the composition further includes a sulfated,galactose-based polysaccharide having an average molecular weightgreater than about 15 kDa. In one embodiment, the galactose-basedpolysaccharide is a carrageenan. In yet another embodiment, thecarrageenan is selected from the group consisting of iota-carrageenan,kappa-carrageenan, and lambda-carrageenan. In a particular embodiment,the carrageenan is iota-carrageenan.

In another embodiment, the galactose-based polysaccharide is derivedfrom an extract isolated from red algae. In yet another embodiment, thegalactose-based polysaccharide is an iota-carrageenan derived from thered algae extract isolated from Gigartina stellata or Chondrus chrispus.In still another embodiment, the galactose-based polysaccharide is akappa-carrageenan derived from the red algae extract isolated fromKappaphycus cottonii. In one embodiment, the red algae is selected fromGigartina stellata, Chondrus chrispus, Kappaphycus cottonii. In anotherembodiment, the galactose-based polysaccharide is in a range of about0.1 to about 10% weight content of the composition. In yet anotherembodiment, the composition includes both a sulfated, galactose-basedpolysaccharide having an average molecular weight greater than about 15kDa and an extract of a blue/green algae.

In yet another embodiment, the ionic aqueous solution includes seawater.In a particular embodiment, the seawater includes undiluted seawater. Ina particular embodiment, the seawater includes (i.e. is diluted with) atleast one of purified, distilled, deionized, and filtered water. Inanother embodiment, the ionic aqueous solution includes saline. In stillyet another embodiment, the ionic aqueous solution includes about 10 toabout 99.9% weight content of the composition.

In another embodiment, the composition has an osmolarity value rangingfrom about 250-350 mOsm/L for isotonic, greater than about 350 mOsm/Lfor hypertonic and less than about 250 mOsm/L for hypotonic. In yetanother embodiment, the composition further includes at least one ofexogenous salts, anti-microbial agents, amino acids, and essential oils.

In another aspect, a use of the composition is provided for themanufacture of a medicament or product for alleviating a respiratorytract and/or respiratory mucosal-related condition. In one embodiment,the composition is formulated for nasal administration. In anotherembodiment, the use is for a respiratory tract and/or respiratorymucosal-related condition selected from the group consisting of acuteand chronic rhinosinusitis, sinusitis, allergic and non-allergicrhinitis, asthma, cystic fibrosis, chronic obstructive pulmonary disease(COPD), and other diseases of the respiratory tract.

In yet another aspect, a composition for nasal administration comprisingan ionic aqueous solution and an extract from brown algae is provided.In one embodiment, the brown algae extract is in a range of about 0.1 toabout 10% weight content of the composition. In another embodiment, thecomposition also includes an extract from a blue/green algae. In aparticular embodiment, the blue/green algae extract is in a range ofabout 0.1 to about 10% weight content of the composition. In yet anotherembodiment, the composition further includes an extract from a redalgae. In one particular embodiment, the red algae extract is in a rangeof about 0.1 to about 10% weight content of the composition. In anotherembodiment, the ionic aqueous solution comprises at least one ofseawater or saline solution. In still another embodiment, thecomposition further includes at least one of exogenous salts,anti-microbial agents, amino acids, and essential oils. In yet anotherembodiment, the composition further includes at least one of acorticosteroid, other steroidal compound, anti-histamine, decongestant,bronchodilator, β2-adrenergic agonist, and antibiotic. In anotherembodiment, the composition further includes at least one plant extract.In another embodiment, the composition further includes at least onehydrating agent.

In still another aspect, pharmaceutical compositions of the hereindescribed compositions are provided.

In another aspect, a pharmaceutical composition for treatment of arespiratory tract and/or respiratory mucosal-related condition includesan ionic aqueous solution, an extract from brown algae and apharmaceutically acceptable carrier or adjuvant. In one embodiment, theionic aqueous solution comprises at least one of seawater or salinesolution. In another embodiment, the respiratory tract and/orrespiratory mucosal-related condition is selected from the groupconsisting of acute and chronic rhinosinusitis, sinusitis, allergic andnon-allergic rhinitis, asthma, cystic fibrosis, chronic obstructivepulmonary disease (COPD), and other diseases of the respiratory tract.In yet another embodiment, the pharmaceutical composition furtherincludes an extract from a blue/green algae. In still yet anotherembodiment, the pharmaceutical composition further includes at least oneof exogenous salts, anti-microbial agents, amino acids, and essentialoils. In yet another embodiment, the composition further includes atleast one of a corticosteroid, other steroidal compound, anti-histamine,decongestant, bronchodilator, β2-adrenergic agonist, and antibiotic.

In yet another aspect, a method is provided for treating a respiratorytract and/or respiratory mucosal-related condition in a subject in needthereof by aerosolizing the pharmaceutical composition described hereinin a nasal passageway of the subject. In one embodiment, the subject isa human subject. In another embodiment, the respiratory tract and/orrespiratory mucosal-related condition is selected from the groupconsisting of acute and chronic rhinosinusitis, sinusitis, allergic andnon-allergic rhinitis, asthma, cystic fibrosis, chronic obstructivepulmonary disease (COPD), and other diseases of the respiratory tract.In a particular embodiment, the respiratory tract and/or respiratorymucosal-related condition is rhinosinusitis.

In still yet another aspect, a packaged device is provided that includesthe pharmaceutical composition described herein optionally together withinstructions for use. In one embodiment, the device is selected from thegroup consisting of aerosol dispenser, pneumatically pressurized device,multi-dose metered dose spray pump, inhaler, pump sprayer, andnebulizer.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the meaning commonly understood by a person skilled in the art towhich this invention performs. As used herein, the following terms havethe meanings ascribed to them below, unless specified otherwise.

By “algae extract” is meant any fraction, or isolated or purifiedmolecule from an algae cell. The extract can include proteins orglycoproteins or nucleic acids. Examples of algae extract include brownalgae extract (e.g., Undaria pinnafitida), blue-green algae extract(e.g., Spirulina platensis), and red algae extract (e.g., Gigartinastellata, Chondrus chrispus, and Kappaphycus cottonii).

By “fucose-based polysaccharide” includes fucan sulfate, fucose-richsulfated polysaccharides, a rhamnose-containing sulfated polysaccharide,a galactofucan and the like. The term “fucoidan” may also be used todescribe the fucose-based polysaccharides from brown seaweeds, as wellas physically, chemically and/or enzymatically derivatized formsthereof.

The term “galactose-based polysaccharide” includes D-galactoses bondedby alpha (1-3) and beta (1-4) bonds, carrageenans (iota-, kappa- andlambda-carrageenan that differ slightly in their structure and degree ofsulfation), linear or branched sulfated galactose-based polysaccharidesand the like. The term “carrageenan” may also be used to describe thegalactose-based polysaccharides extracted from red edible seaweed, aswell as physically, chemically and/or enzymatically derivatized formsthereof.

By “respiratory tract and/or respiratory mucosal-related conditions,”are meant any conditions or disorders with abnormal mucus production,secretion or clearance or inflammation (bacterial, viral, allergic orautoimmune) of the nasal, bronchial and pulmonary mucosa. Examples ofdiseases include acute rhinosinusitis, recurrent acute rhinosinusitis,subacute rhinosinusitis, chronic rhinosinusitis, acute exacerbation ofchronic rhinosinusitis, sinusitis, allergic and non-allergic rhinitis,asthma, cystic fibrosis, chronic obstructive pulmonary disease (COPD),and other diseases of the upper and lower respiratory tract.

By “agent” is meant any small molecule compound or mixes thereofsynthesized chemically or derived from natural sources (e.g. herbalextracts), antibody, nucleic acid molecule, or polypeptide, or fragmentsthereof.

By “ameliorate” is meant decrease, suppress, attenuate, diminish,arrest, or stabilize the development or progression of a disease.

In this disclosure, “comprises,” “comprising,” “containing” and “having”and the like have the meaning ascribed to them in U.S. Patent law andcan mean “includes,” “including,” and the like; “consisting essentiallyof” or “consists essentially” likewise has the meaning ascribed in U.S.Patent law and the term is open-ended, allowing for the presence of morethan that which is recited so long as basic or novel characteristics ofthat which is recited is not changed by the presence of more than thatwhich is recited, but excludes prior art embodiments.

By “effective amount” is meant the amount of a composition of theinvention required to ameliorate the symptoms of rhinosinusitis of anuntreated patient. The effective amount of active compound(s) used topractice the present invention for therapeutic treatment ofrhinosinusitis varies depending upon the manner of administration, theage, body weight, and general health of the subject. Ultimately, theattending physician or veterinarian will decide the appropriate amountand dosage regimen. Such amount is referred to as an “effective” amount.

A “fraction” of an organism, such as a species of algae, is any isolatedor purified molecule or complex of molecules. A fraction can be obtainedby any method known in the art by which molecules are isolated orpurified from a cell, such as by extraction or ultracentrifugation. Inparticular examples, a fraction of algae is produced by extraction ofthe algae cell followed by further isolation of molecules of aparticular molecular weight range or size. In particular examples, afraction is in liquid form. In other examples, a fraction is in a solidform, such as a dried form. In still other examples, a fraction can beformulated as an aerosolized particulate.

The terms “isolated,” “purified,” or “biologically pure” refer tomaterial that is free to varying degrees from components which normallyaccompany it as found in its native state. “Isolate” denotes a degree ofseparation from original source or surroundings. “Purify” denotes adegree of separation that is higher than isolation. A “purified” or“biologically pure” polysaccharide is sufficiently free of othermaterials such that any impurities do not materially affect thebiological properties of the polysaccharide or cause other adverseconsequences. That is, a polysaccharide of this invention is purified ifit is substantially free of cellular or other materials when isolated orchemically synthesized. Purity and homogeneity are typically determinedusing analytical chemistry techniques, for example, electrophoresis orthin layer chromatography. The term “purified” can denote that amaterial that is enriched in polysaccharides by removing proteins and/orDNA.

As used herein, “obtaining” as in “obtaining an agent” includessynthesizing, purchasing, or otherwise acquiring the agent.

As used herein, the terms “prevent,” “preventing,” “prevention,”“prophylactic treatment” and the like refer to reducing the probabilityof developing respiratory tract and/or respiratory mucosa-relatedconditions, e.g., rhinosinusitis or inflammatory condition, in asubject, who does not have, but is at risk of or susceptible todeveloping such conditions.

By “reduces” is meant a negative alteration of at least 10%, 25%, 50%,75%, or 100%.

By “subject” is meant a mammal, including, but not limited to, a humanor non-human mammal, such as a bovine, equine, canine, ovine, or feline.

Ranges provided herein are understood to be shorthand for all of thevalues within the range. For example, a range of 1 to 50 is understoodto include any number, combination of numbers, or sub-range from thegroup consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

As used herein, the terms “treat,” treating,” “treatment,” and the likerefer to reducing or ameliorating a disorder and/or symptoms associatedtherewith. It will be appreciated that, although not precluded, treatinga disorder or condition does not require that the disorder, condition orsymptoms associated therewith be completely eliminated.

Unless specifically stated or obvious from context, as used herein, theterm “or” is understood to be inclusive. Unless specifically stated orobvious from context, as used herein, the terms “a”, “an”, and “the” areunderstood to be singular or plural.

Unless specifically stated or obvious from context, as used herein, theterm “about” is understood as within a range of normal tolerance in theart, for example within 2 standard deviations of the mean. About can beunderstood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%,0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear fromcontext, all numerical values provided herein are modified by the termabout.

As used herein “% weight content” means % w/w. All percentage valuesmentioned herein are % w/w unless indicated otherwise.

As used herein, a “substantially similar composition” in relation to areference formulation means a formulation having the same components inthe same concentration as in the reference, or wherein the concentrationof any of the components differs relative to the reference in animmaterial way.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable or aspect herein includes that embodiment as any singleembodiment or in combination with any other embodiments or portionsthereof.

Any compositions or methods provided herein can be combined with one ormore of any of the other compositions and methods provided herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 presents data obtained after testing Formulations 1-4 and variouscontrols for IL-8 secretion in a three-dimensional reconstituted humanairway epithelia model utilizing cells isolated from normal human donors(see Example 1).

FIG. 2 presents data obtained after testing Formulation 1 and variouscontrols for mucociliary clearance (Plate A), IL-8 secretion (Plate B)and cilia beating frequency (Plate C) in a three-dimensionalreconstituted human airway epithelia model utilizing cells isolated froma human allergic donor (see Example 2).

DETAILED DESCRIPTION OF THE INVENTION

This invention provides seawater-based compositions. Compositions withingredients that are widely abundant, easily isolated by naturallygrowing organisms, intrinsically compatible with seawater and/or salinesolutions of all types of osmolarities and generally recognized as safe,are particularly appealing for the development of new solutions withimproved efficacy and negligible side effects. The compositionsdescribed herein are believed to satisfy all these conditions.

The compositions can be useful as nasal passage washes to aid in theresorption of edema of the nasal mucosa, and/or for the treatment ofrespiratory tract and/or respiratory mucosal-related conditions, such aschronic rhinosinusitis, sinusitis, allergic rhinitis, nasal polyps, andother nasal conditions and/or inflammatory conditions of nasal,bronchial and respiratory mucosa. The composition includes an ionicaqueous solution and algae-derived constituents, such as sulfatedpolysaccharides having an average molecular weight greater than 4 kDaand comprising L-fucose and sulfate ester groups. The sulfatedpolysaccharides may be linear or branched.

The composition can be applied in the form of a nasal spray (oralternatively in an aerosolized/nebulized form for inhalation) topatients with various inflammatory and other conditions affecting thenasal, bronchial and pulmonary mucosa caused by various agents (e.g.substances of bacterial origin, viruses, bacteria, irritants, allergensetc). Some nonlimiting examples of inflammatory conditions include:acute and chronic rhinosinusitis, allergic and non-allergic rhinitis,asthma, cystic fibrosis, chronic obstructive pulmonary disease (COPD),other diseases of the respiratory tract, and others.

Compositions

Described herein are compositions, such as seawater-based compositions,intended for administration to and through the respiratory tract, e.g.,via nasal administration. The compositions can include a mixture ofpreviously filtered, purified and sterilized seawater and an algae-basedconstituent. In one aspect, the invention provides a composition with anionic aqueous solution and an algal extract, such as brown algae.Compositions of seawater including ionic aqueous solutions, such assaline or seawater solutions, and algae-derived constituents aredescribed further below.

Ionic Aqueous Solutions

Compositions of the invention can include an ionic aqueous solution thatis filtered and/or sterilized. Ionic aqueous solutions can be classifiedinto two different categories: saline solutions and seawater solutions.

Seawater sterilization has been previously described. EP 0909184 alsodescribes relative percentages of magnesium, calcium, potassium, zincand copper presented in seawater collected in Cancale Bay. Additionalsites in the same area (Saint Malo) can also be used as the sources ofseawater, see FR2915389. The seawater collected can be diluted todifferent extents with purified/filtered/deionized water or used forselective electrodialysis to alter the different ionic concentrations,see also WO/2012/110665.

Saline solutions can refer to aqueous solutions comprising variousconcentrations of sodium chloride (NaCl) including: (1) normal orisotonic saline solutions: solutions containing from about 0.9% w/v ofNaCl and an osmolarity of about 300 mOsm/L; (2) hypotonic saline,solutions containing NaCl of less than about 0.9% NaCl and less thanabout 300 mOsm/L, see for example U.S. Pat. No. 8,337,906; and (3)hypertonic saline solutions containing NaCl of greater than about 0.9%NaCl and greater than about 300 mOsm/L. For example, solutions of 3% and5% NaCl would have osmolarities of about 1000 mOsm/L and about 1700mOsm/L, respectively.

The saline solutions can include additional salts, such as thenon-limiting examples of, Dead Sea salts and anti-oxidants, asexemplified by U.S. Pat. No. 7,541,052, see also U.S. Pat. No.6,528,081. Optional additional ingredients can could include zinc, asexemplified by U.S. Pat. No. 5,622,724, copper, manganese, magnesium andother metal ions, anti-microbial agents, essential oils, etc, see alsoWO/2003/015746 and WO/2012/110665. These solutions may have the pHadjusted, i.e. to physiological levels (pH 7-8) using a variety ofbuffers. Saline solutions may also include hypertonic (U.S. Pat. No.7,541,052) or hypotonic saline solutions.

Seawater solutions can refer to three types of solutions: (1) undilutedseawater; (2) diluted seawater; and (3) ion content/osmolarity alteredseawater.

Undiluted seawater contains a multitude of ingredients, see Table 1below; these include various salts, minerals and trace elements. It isnoted that the ionic content/strength of seawater is measured in 2 ways:either as sodium chloride equivalents or via direct measurements ofosmolarity (e.g., about 1000 mOsm/L).

In the former case, NaCl equivalents are calculated using methods, suchas the Volhard method. In this case, the method precipitates allchlorides in seawater (“halides” e.g. Na, K, Ca, Mg salts with CI) andconverts them into a percentage of NaCl. Accordingly, undiluted seawaterhas about 3.1-3.8% w/v NaCl (depending on the origin of seawater).

Regarding the osmolarity measurements of pure seawater, it is noted thatthe contribution of different ions (in mOsm) in the about 1000 mOsm/Lvalue are: Na: 459, K: 10, Ca:10, Mg:53, Cl:538. Seawater used in thiscase is typically filtered and/or sterilized to remove/eliminatemicroorganisms before use.

Seawater can be diluted to different degrees withpurified/distilled/deionized/filtered water. Depending on the dilutingfactor, solutions can be rendered hypotonic, isotonic or increasinglyless hypertonic in comparison to undiluted seawater. For example, a 7:3dilution is described in EP 0908184, which is incorporated by referencein its entirety. This results in a hypertonic solution comprising about2.3-2.5% NaCl (equivalents). Additional dilutions can also be done togive solutions with about 0.9, 2, 2.6 or 3% NaCl, see for example U.S.Pat. No. 4,581,226, (also Tables 1 and 3). It is obvious that theosmolarity of seawater is also changing according to dilution.

Ion content/osmolarity of seawater can also be artificially altered bychemicals means, e.g. electrodialysis to specifically reduce the contentof ions to the desired extent (e.g., to about 0.9% NaCl rendering thesolution iso-osmotic) or to about 2.2% (hyperosmotic). Typical solutionsof this kind are described in EP 1091747 or US2010/0151044 (iso-osmotic)or WO/2008/037938 and WO/2012/110665 (hyperosmotic).

The difference between diluted and electrodialyzed seawater solutions isthat apart from the differential percentage of NaCl, seawateringredients are diluted in the former case whereas in the latter casethey remain as they are in pure seawater. The relevance of thisinformation is explained below, see also Tables 2 and 3 below for adirect comparison between ion contents in both methods. It is noted thatas pure seawater has various % s of the different ingredients to startwith, so there is some variability in the preparation of solutions ineach case.

TABLE 1 Composition of seawater from “Handbook of Chemistry and Physics”63^(rd) edition 1982-1983, CRC Press, see also Table A ofUS2010/0151044. Quantity Element (ppm) Element Quantity (ppm) Cl 18,980Pb 0.004-0.005 Na 10,561 Se 0.004 Mg 1,272 Sn 0.003 S 884 Cs 0.002(approximately) Ca 400 U 0.00015-0.0016  K 380 Mo 0.0003-0.002  Br 65 Ga0.0005 C (inorganic) 28 Ni 0.0001-0.0005 Sr 13 Th <0.0005 (SiO2)0.01-7.0 Ce 0.0004 B 4.6 V 0.0003 C (organic)  1.2-3.0 La 0.0003 Al0.16-1.9 Y 0.0003 F 1.4 Hg 0.00003 N (nitrate) 0.001-0.7  Ag0.00015-0.0003  N (organic nitrogen) 0.03-0.2 Bi 0.0002 Rb 0.2 Co 0.0001Li 0.1 Sc 0.00004 P (phosphate) >0.001-0.10  Au 0.000004-0.000008 Ba0.05 Fe (in true <10-9  solution) I 0.05 Ra 2.10-11 = 3.10--10 N(nitrite) 0.0001-0.05  Ge Present N (ammoniac) >0.005-0.05  Ti PresentAs (arsenic)  0.003-0.024 W Present Fe 0.002-0.02 Cd Present in marineorganisms P (organic 0.016 Cr Present in marine phosphorus) organisms Zn 0.005-0.014 Tl Present in marine organisms Cu 0.001-0.09 Sb Present inmarine organisms Mn 0.001-0.01 Zr Present in marine organisms Pt Presentin marine organisms

TABLE 2 Ion content/osmolarity altered by seawater dilution tospecifically reduce the content of ions to the desired extent PureSeawater* Isotonic Hypertonic (mg/L)** % Sodium NaCl*** 3.50% 0.90%2.00% 2.30% 2.60% 3.00% Chloride Sodium Na+ 10,500 2,700 6,000 6,9007,800 9,000 Chloride Cl− 19,000 4,886 10,857 12,486 14,114 16,286Potassium K+ 390 100 223 256 290 334 Calcium Ca++ 410 105 234 269 305351 Magnesium Mg++ 1,350 347 771 887 1,003 1,157 Sulfate SO4 −− 2,700694 1,543 1,774 2,006 2,314 *pure seawater values are taken fromLangmuir (1997): Aqueous environmental geochemistry: Upper Saddle River,Prentice Hall, Inc., 600p **depicted values will vary 5-10% depending onorigin of seawater ***calculated as NaCl equivalents (Volhard method)

TABLE 3 Ion content/osmolarity altered by electrodialysis tospecifically reduce the content of ions to the desired extent Pure Iso-Hyper- Seawater Osmotic Osmotic (mg/L) % Sodium NaCl 3.50% 0.90% 2.20%Chloride Sodium Na+ 10,500 2100-2600 5500-7500 Chloride Cl− 19,0005400-6300 10000-13000 Potassium K+ 390 44-62 100-250 Calcium Ca++ 410280-390 300-450 Magnesium Mg++ 1,350 1100-1500 1100-1500 Sulfate SO4−−2,700 2,700 2,700

Algae Extracts

Compositions of the invention can also include algae extracts, such assulfated polysaccharides obtained from algae extracts. Algae extractscan be obtained from three different algae: (1) brown algae; (2) blueand green algae; (3) and red algae.

Brown Algae Extracts

Also provided are compositions that include extracts from brown algae.Brown algae extracts and some marine invertebrates (such as sea urchinsand sea cucumbers) are rich in sulfated polysaccharides. Sulfatedpolysaccharides obtained from brown algae extracts can have an averagemolecular weight greater than about 4 kDa. The sulfated polysaccharidescan be linear or branched. The sulfated polysaccharides includefucose-based polysaccharides with L-fucose and sulfate ester groups,such as fucoidan. Fucoidans include those derived from brown algae, suchas Undaria pinnafitida, or synthetic fucoidan polysaccharides, such as ahigh molecular weight fucoidan (HMWF) fraction having an averagemolecular weight ranging from about 1 to 2 MDa (e.g. Kraeber, Germany)and a low molecular weight fucoidan (LMWF) fraction having an averagemolecular weight of about 8.2 kDa, are useful.

Particular brown algae include species of Ascoseira, Cutleria,Microzonia, Zanardinia, Arthrocladia, Desmarestia, Himantothallus,Phaeurusm, Dictyopteris, Dictyota, Dilophus, Distromium, Glossophora,Homoeostrichus, Lobophora, Lobospira, Newhousia, Pachydictyon, Padina,Spatoglossum, Stypopodium, Taonia, Zonaria, Scoresbyella,Choristocarpus, Discosporangium, Acinetospora, Feldmannia, Geminocarpus,Hincksia, Pogotrichum, Pylaiella, Adenocystis, Caepidium, Utriculidium,Acrothrix, Ascoseirophila, Asperococcus, Austrofilum, Chordaria,Cladosiphon, Corycus, Delamarea, Dictyosiphon, Elachista, Eudesme,Giraudia, Gononema, Halothrix, Haplogia, Hecatonema, Heterosaundersella,Hummia, Isthmoplea, Laminariocolax, Laminarionema, Leathesia,Leptonematella, Litosiphon, Microspongium, Mikrosyphar, Myelophycus,Myriogloea, Myrionema, Myriotrichia, Papenfussiella, Petrospongium,Pleurocladia, Polytretus, Proselachista, Protectocarpus, Punctaria,Sauvageaugloia, Soranthera, Sorocarpus, Spermatochnus, Sphaerotrichia,Stictyosiphon, Streblonema, Striaria, Stschapovia, Tinocladia,Chordariopsis, Asterocladon, Ectocarpus, Kuckuckia, Mesospora,Asterotrichia, Bachelotia, Bifurcariopsis, Durvillaea, Ascophyllum,Fucus, Hesperophycus, Pelvetia, Pelvetiopsis, Silvetia, Xiphosphora,Himanthalia, Hormosira, Notheia, Anthophycus, Axillariella, Bifurcaria,Bifurcariopsis, Carpoglossum, Caulocystis, Coccophora, Cystophora,Cystoseira, Halidrys, Hizikia, Hormophysa, Myagropsis, Myogropsis,Myriodesma, Sargassum, Turbinaria, Cystophaera, Marginariella,Phyllospora, Seirococcus, Ishige, Akkesiphycus, Alaria, Aureophycus,Druehlia, Eualaria, Hirome, Lessoniopsis, Pleurophycus, Pterygophora,Undaria, Undariella, Undariopsis, Chorda, Agarum, Costaria,Dictyoneurum, Thalassiophyllum, Arthrothamnus, Costularia, Cymathere,Feditia, Gigantea, Laminaria, Macrocystis, Nereocystis, Pelagophycus,Pelagophycus, Macrocystis, Phycocastanum, Phyllariella, Polyschidea,Postelsia, Pseudolessonia, Saccharina, Streptophyllopsis, EckIonia,Eckloniopsis, Egregia, Eisenia, Lessonia, Pseudochorda, Nemoderma,Onslowia, Verosphacella, Neoralfsia, Basispora, Hapalospongidion,Jonssonia, Lithoderma, Myrionemopsis, Petroderma, Porterinema,Pseudolithoderma, Ralfsia, Chnoospora, Colpomenia, Hydroclathrus,Petalonia, Rosenvingea, Scytosiphon, Bodanella, Coelocladia,Heribaudiella, Phaeostroma, Asteronema, Scytothamnus, Stereocladon,Splachnidium, Cladostephus, Sphacelaria, Sphacella, Alethocladus,Halopteris, Stypocaulon, Austronereia, Bellotia, Carpomitra,Encyothalia, Nereia, Perisporochnus, Perithalia, Sporochnema,Sporochnus, Tomaculopsis, Syringoderma, Halosiphon, Masonophycus,Phyllariopsis, Saccorhiza, Stschapovia, Haplospora, Phaeosiphoniella,Tilopteris, Neolepioneuma, Analipus and Phaeostrophion.

Other sources include some green seaweeds and echinoderms such as seaurchins and sea cucumbers. Examples of green seaweed include Ulva sp,Enteromorpha sp, Codium sp, Caulerpa sp and Halimala sp.

The sulfated polysaccharides that include fucose-based polysaccharides,can include extracts derived from Fucus vesiculosus and contain mostlyfucose, sulfate, ash, and traces of aminoglucose. Sulfatedpolysaccharides from brown algae F. evanescens C. Ag, F. distichus andF. serratus L. include mostly fucose, sulfate and acetate. Fucoidansinclude F-fucoidan, mainly composed of sulfated esters of fucose, andU-fucoidan, composed of about 20% of glucuronic acid. Some sulfatedpolysaccharides include alternating 3- and 4-linked α-L-fucopyranose2-sulfate residues and others can include disaccharide repeating units.In addition to linear polysaccharides, highly branched structures areincluded in extract derived from E. kurome or Chorda filum.

The sulfated polysaccharides for use in the method and compositionsdescribed herein may include an extract from the whole plant or any partof the plant, such as the leaves, stem, spores, or a combinationthereof. The starting material may be fresh, frozen or dried material.After filtration or dialysis, the extract may be used as a liquid ordried. Typically, such an extract includes from at least about 1% w/vfucose-based polysaccharide to about 100% w/v.

The sulfated polysaccharides are generally from about 4 kDa to about 5MDa including about 5, 10, 15, 20, 30, 50, 70, 90, 100, 300, 500, 700,900, 1000, 2000, 3000, 4000 and 5000 kDa or an average molecular weightin between these particular sizes.

The sulfated polysaccharides also include fucoidans described in patentapplication WO/2011/100805, see for example relevant information thatcan be used on MW (greater than about 4 kDa, preparations comprisingvarious fucoidan contents e.g. about 10-95% or more), species that canbe used for isolation, specifications of fucoidans used etc. Note thatfucoidans used in the Examples are derived from Undaria pinnafitida(aqueous) extracts and comprise greater than about 85% of fucoidans.

In one embodiment, compositions include about 0.1% to about 10% sulfatedpolysaccharide or fucoidan weight content of the composition. The rangeof sulfated polysaccharide or fucoidan in the composition can be fromabout 0.1% to about 20%, about 0.1% to about 10%, about 0.1% to about5%, and about 0.1% to about 1% weight content of the composition. Inother embodiments, the composition includes at least about 0.1%, 0.2%,0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 4%,5%, 6%, 7%, 8%, 9%, 10%, or more % sulfated polysaccharide or fucoidanweight content.

Blue-Green Algae Extracts

The compositions of the invention may also include extracts fromblue-green algae. Blue-green algae extracts are rich in protein,vitamins, minerals, and carotenoids, antioxidants that can help protectcells from damage. Extracts contain nutrients, including B complexvitamins, beta-carotene, vitamin E, manganese, zinc, copper, iron,selenium, and gamma linolenic acid (an essential fatty acid). Blue-greenalgae, such as Aphanizomenon flos aquae (AFA) or Spirulina (Arthrospira)have up about 62% amino acids in their extracts. Because extracts are arich source of protein and other nutrients, they have been used innutritional supplements.

Blue-green algae, such as AFA or Spirulina (Arthrospira), can beisolated from any source. The source can be a naturally occurring sourcethat rich in blue-green algae. The source can also be a man-made source,such as an artificial lake or water source. The source can also bebioreactors or fermentors. The source can be an environment produced togrow and harvest blue-green algae commercially.

The blue-green algae extracts can include an extract from the whole algaor any part of the alga. The extract can be fractionated. The extract ofblue-green algae can include any fraction, or isolated or purifiedmolecule from a blue-green alga cell. The blue-green algae aredisrupted, an inorganic or organic solvent is added, and extract (ormolecules) is collected. Specific, non-limiting examples of extract areisolated using high performance liquid chromatography, thin layerchromatography, affinity column, magnetic beads or distillation. In oneembodiment, fractionation is used to isolate the extract based on themolecular weight or the hydrophobicity of the molecules of theblue-green algae. For example, the extract can include those moleculesof less than about 50 kDa, and not 50 kDa or greater; less than about 10kDa, and not 10 kDa or greater; or less than about 5 kDa, and not 5 kDaor greater.

An extract of blue-green algae, such as a dried form, include about 0.5%to about 5% blue-green algae extract weight content of the composition.The range of blue-green algae extract in the composition can be fromabout 0.1% to about 20%, about 0.5% to about 20%, about 0.5% to about10%, about 0.5% to about 5%, and about 0.5% to about 1% weight contentof the composition. In other embodiments, the composition includes atleast about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%,1.5%, 2%, 2.5%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or more blue-green algaeextract weight content. In an exemplary embodiment, blue-green algaeextract is in range of about 0.1% to about 10% of the composition.

Red Algae Extracts

The compositions provided may include extracts from red algae. Red algaecontain sulfated polysaccharides with D-galactoses bonded alternativelyby alpha (1-3) and beta (1-4) bonds. These galactose-basedpolysaccharides are distinguished by the presence or not of a 3,6anhydrous bridge on the alpha bonded (1-3) galactose residue and bytheir level of sulfation. Carrageenan is a generic term for the linearor branched sulfated galactose-based polysaccharides extracted from redalgae (rhodophyceae or red seaweed). Examples in this case are describedin WO/2009/027057.

Methods of galactose-based polysaccharide or carrageenan extraction andproduction can be derived from wild-grown red algae, likecarrageenan-containing plants such as Gigartina radula, Gigartinaskottsbergii, Gigartina chamissoi, Gigartina stellata, Iridaea cordata,Chondrus chrispus and Sarcothalia crispate seeded along nylon ropes andharvested in massive aqua-culture farming operations or obtained fromsynthetic carrageenan production. Isolation of the galactose-basedpolysaccharides from the red algae is conducted in accordance withstandard techniques, as described in U.S. Pat. No. 3,094,517 to Stanleyet al. The carrageenan is extracted from the cleaned algae while alsobeing subjected to alkali modification by placing the algae in asolution made slightly alkaline by the addition of a low concentrationof alkali salt (i.e., a pH of the solution is raised to a range of,e.g., 9-10) and then heating this solution to a temperature of around80° C. for a period of time of about 20 minutes to as long as two hours.

There exist more than 10 structurally different carrageenans, theirnature depending on the algae genus from which they are extracted. Thethree main types are iota-, kappa- and lambda-carrageenan, which differslightly in their structure and degree of sulfation. Iota-carrageenan isa soft-gel forming sulfated galactose-based polysaccharide predominantlyextracted from red seaweed, Gigartina stellata and Chondrus crispus.Kappa-carrageenan yields strong, rigid gels and is predominantlyproduced from Kappaphycus cottonii. Lambda-carrageenan, which is themost common form, is frequently used to thicken dairy products.

The galactose-based polysaccharides can have a molecular weight rangingfrom about 15 kDa to 5 MDa, and fractions having average molecularweights of more than 50 kDa, and especially fractions having averagemolecular weights in the range of from 50 kDa to 3 MDa. Thegalactose-based polysaccharides also include carrageenans described inpatent application WO/2009/027057, see for example for relevantinformation.

In one embodiment, compositions include about 0.1% to about 10%galactose-based polysaccharide or carrageenan weight content of thecomposition. The range of galactose-based polysaccharide or carrageenanin the composition can be from about 0.1% to about 20%, about 0.1% toabout 10%, about 0.1% to about 5%, and about 0.1% to about 1% weightcontent of the composition. In other embodiments, the compositionincludes at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or moregalactose-based polysaccharide or carrageenan weight content.

Algae Extraction

As provided herein, algae extracts can be prepared from fresh,dehydrated, or preserved algae or marine cells. The algae can beextracted in any aqueous solution, such as with water or a suitablebuffered salt solution, or other solvents, such as ethanol and methanol,and DMSO. The extract may also be produced by any other suitable method.Following extraction, the extracts are dried, as described above. Thealgae extracts can be used directly, or can be stored as liquid, frozenliquid, dehydrated, freeze-dried, vacuum dried, drum dried cells andspray-dried cells.

In one embodiment, the extracts are filtered to separate fractions ofdifferent sizes. The filtration can be performed using ultra filtrationfilters having different molecular weight cut-offs, such as a filterwith a 10 MDa cut-off that allows collection of molecules of less thanabout 10 MDa, and excludes molecules of about 10 MDa or greater. In thiscontext, “about” refers to a difference of 5 kDa. In other examples, themolecular weight cut-off filter allows collection of molecules that areless than 5 MDa, and excludes molecules of 5 MDa or greater; less than 1MDa, and excludes molecules of 1 MDa or greater; less than 500 kDa, andexcludes molecules of 500 kDa or greater; less than 300 kDa, andexcludes molecules of 300 kDa or greater; less than 100 kDa, andexcludes molecules of 100 kDa or greater; or less than 50 kDa, andexcludes molecules of 50 kDa or greater.

Other Components

The compositions provided herein can include one or more additionalcomponents. Non-limiting examples of additional components/agentsoptionally included in the composition include cooling agents such asmenthol, warming agents, flavoring agents, salivating agents, teaextract, exogenous salts (e.g. Dead Sea salts), anti-septic andanti-bacterial agents, amino acids, essential oils, vitamin(s) (e.g.,Vitamin A, Vitamin C, Vitamin B, and/or Vitamin D), carotenoid,rosemary, rosemary extract, caffeic acid, coffee extract, tumericextract, curcumin, blueberry extract, grape seed extract, rosemaricacid, antioxidant, enzyme, prebiotic, probiotic, and rographis extract,1-tryptophan, Allium sativum, herbal remedies, supplements, naturalingredients, minerals, energy boosting ingredients, sleep aids, immunesystem boosting agents, colorant, preservative, fragrance, fruitextract, others agents, and combinations thereof. Further potentialadditional components include hydrating agents.

Essential oils can be added to represent characteristically scented,essential herbal active agents, which can be disinfecting or impartantimicrobial action. Lemon oil, eucalyptus oil, balm oil, mint oil,camphor, aniseed oil, rosemary oil, spearmint oil, peppermint oil,thymus oil and sage oil can be used as essential oils. The concentrationof the essential oils can be, approximately, 0.01 mg to 50 mg, inparticular 0.05 mg to 15 mg per 10 ml, whereby 0.01 mg to 10 mg, inparticular, 0.01 mg to 5 mg is lemon oil; 0.01 mg to 10 mg, inparticular, 0.01 mg to 5 mg is eucalyptus oil; 0.01 mg to 5 mg, inparticular, 0.01 mg to 0.3 mg is balm oil; 0.01 mg to 10 mg, inparticular, 0.01 mg to 3 mg is mint oil; 0.01 mg to 10 mg, inparticular, 0.01 mg to 3 mg is campho; 0.001 mg to 5 mg, in particular,0.001 mg to 0.5 mg is aniseed oil; 0.001 mg to 5 mg, in particular,0.001 mg to 0.05 mg is rosemary oil; 0.01 mg to 10 mg, in particular,0.01 mg to 5 mg is spearmint oil; 0.01 mg to 10 mg, in particular, 0.01mg to 5 mg is peppermint oil; 0.01 mg to 10 mg, in particular, 0.01 mgto 5 mg is thymus oil; and 0.001 mg to 5 mg, in particular, 0.001 mg to0.5 mg is sage oil per 10 ml of the composition.

The composition can include a pharmaceutically acceptable carrier, e.g.,one or more solvents, dispersion media, coatings, antimicrobial agents,isotonic and absorption delaying agents, and the like, compatible withadministration to a mammal, such as a human. Any carrier compatible withthe excipient(s) and therapeutic agent(s) is suitable for use.Supplementary active compounds may also be incorporated into thecompositions.

Any antimicrobial agent known in the art can be used in the compositionsat concentrations generally used for such agents. Antimicrobial agentsinclude antibacterials, antifungals, and antivirals. Exemplaryantibiotics (i.e., antibacterial agents) include the penicillins (e.g.,penicillin G, ampicillin, methicillin, oxacillin, and amoxicillin), thecephalosporins (e.g., cefadroxil, ceforanid, cefotaxime, andceftriaxone), the tetracyclines (e.g., doxycycline, minocycline, andtetracycline), the am inoglycosides (e.g., am ikacin, gentamycin,kanamycin, neomycin, streptomycin, and tobramycin), the macrolides(e.g., azithromycin, clarithromycin, and erythromycin), thefluoroquinolones (e.g., ciprofloxacin, lomefloxacin, levofloxacin andnorfloxacin), and other antibiotics including chloramphenicol,clindamycin, cycloserine, isoniazid, rifampin, sulfamethoxazole withtrimethoprim, cefuroxime, cefpodoxime, and vancomycin. Antiviral agentsare substances capable of inhibiting the replication of viruses.Examples of anti-viral agents include1,-D-ribofuranosyl-1,2,4-triazole-3 carboxamide, 9-2-hydroxy-ethoxymethylguanine, adamantanamine, 5-iodo-2′-deoxyuridine,trifluorothymidine, interferon, adenine arabinoside, proteaseinhibitors, thymidine kinase inhibitors, sugar or glycoprotein synthesisinhibitors, structural protein synthesis inhibitors, attachment andadsorption inhibitors, and nucleoside analogues such as acyclovir,penciclovir, valacyclovir, and ganciclovir. Antifungal agents includeboth fungicidal and fungistatic agents such as, for example, benzoicacid, undecylenic alkanolamide, ciclopirox olamine, polyenes,imidazoles, allylamine, thicarbamates, amphotericin B, butylparaben,clindamycin, econaxole, fluconazole, flucytosine, griseofulvin,nystatin, and ketoconazole.

The composition can also be formulated in combination with other agentstypically used to relieve symptoms of sinusitis, hay fever, nasalcongestion, allergic rhinitis, and/or non-allergic (perennial) rhinitis.Examples of other agents can include but are not limited tocorticosteroids, other steroidal compounds, anti-histamines,decongestants, bronchodilators, β2-adrenergic agonists, andantimicrobial agent. In one embodiment, the composition further includesat least one of a corticosteroid, other steroidal compound,anti-histamine, decongestant, bronchodilator, β2-adrenergic agonist, andantimicrobial agent.

Corticosteroids and other steroidal compounds can be used with thedescribed composition to reduce inflammation and histamine production inthe nasal passages, thereby relieving nasal congestion, runny nose,sneezing, sinus pain, headaches etc. Examples of corticosteroids andother steroidal compounds can include alcometasone, beclomethasone,betamethasone, budesonide, ciclesonide, clobetasol, deflazacort,diflucortolone, desoxymethasone, dexamethasone, fludrocortisone,flunisolide, fluocinolone, fluometholone, fluticasone, hydrocortisone,mometasone furoate, nandrolone decanoate, rimexolone,methylprednisolone, prednisolone and triamcinolone acetonide. Exemplaryexamples of corticosteroids and other steroidal compounds can includemometasone, budesonide, fluticasone, flunisolide, beclomethasone,triamcinolone, and ciclesonide.

Antihistamines are one of the primary medicaments employed to treatallergic rhinitis. Antihistamines are helpful to control sneezing,itching, and rhinorrhea, as well as associated ocular symptoms, but areineffective in relieving nasal blockage. Antihistamines compete withhistamine for binding to H1 receptors and thereby prevent the action ofhistamine which includes bronchospasm, edema, increased mucus secretionand itching. The antihistamines used today can be given orally ortopically (intranasally). Examples of useful antihistamines that may beused with the described composition can include acrivastine, azatadine,azelastine, astemizole, brompheniramine, carbinoxamine, cetirizine,chloropheniramine, clemastine, cyproheptadine, dexchloropheniramine,diphenhydramine, doxylamine, ebastine, hydroxyzine, ketotifen,levocabastine, lodoxamide, loratadine, meclizine, mequitazine,methdilazine, oxatomide, phenindamine, promethazine, pyrilamine,setastine, rocastine, tazifylline, temelastine, terfenadine,trimeprazine, tripelennamine, and triprolidine. Exemplary examples ofanti-histamines can include astemizole, Azatadine, azelastine,cetirizine, ebastine, ketotifen levocabastine, loratadine, lodoxamide,levocabastine, mequitazine, oxatomide, setastine, tazifylline,temelastine, and terfenadine.

Nasal decongestants include compounds that relieve nasal congestion bynarrowing (constricting) the blood vessels and reducing blood flow,swelling and mucous formation. The vast majority of decongestants actvia enhancing norepinephrine (noradrenaline) and epinephrine(adrenaline) or adrenergic activity by stimulating the α-adrenergicreceptors. Decongestants can be oral (e.g. tablets) in nasal sprays ordrops. Examples of decongestants can include ephedrine,levo-methamphetamine, naphazoline, oxymetazolin, phenylephrine,phenylpropanolamine, propylhexedrine, pseudoephedrine, synephrin,tetrahydrozoline, tramazoline, and xylometazoline.

Bronchodilators are substances that dilate the bronchi and bronchioles,thereby decreasing resistance in the respiratory airway and increasingairflow to the lungs. They are most useful in obstructive lung diseases,of which asthma and chronic obstructive pulmonary disease are the mostcommon conditions. Classes within this category of compounds includeβ2-adrenergic agonists and anticholinergics. β2-adrenergic agonists,also known as β2-adrenergic receptor agonists, are a class of drugs thatact on the β2-adrenergic receptor, thereby causing smooth musclerelaxation, resulting in dilation of bronchial passages. Examples oflong-acting β2-agonists (LABAs) include formoterol, salmeterol and saltsthereof, such as formoterol fumarate and salmeterol xinafoate. Examplesof short-acting β2-agonists include salbutamol, terbutaline and saltsthereof such as salbutamol sulfate. Examples of anticolinergics includeagents used in COPD and/or asthma such as tiotropium and ipratropiumbromide.

Plant extracts that can be used in the composition include in anon-limiting manner extracts isolated by various means (e.g. viaextraction with aqueous or organic solvents) by different parts of theplant (e.g. root, fruits, leaves etc) such as rosemary extract, coffeeextract, tumeric extract, blueberry extract, grape seed extract,andrographis extract, Allium sativum extract, Thyme extract (e.g,extract of Thymus vulgaris), Centella asiatica extract or extractstypically used in cosmetic and pharmaceutical industry such as thoseincluded in Pharmacopoeia, WHO (World Health Organization) monographs orin the compendium PDR for Herbal Medicines 4th Edition (ThomsonHealthcare). Typical concentrations of plant extracts that could be usedin the composition range between 0.001% to 5%, most preferably between0.05-1% in the composition.

Herbal remedies and other natural ingredients (e.g. carotenoids, caffeicacid, rosemaric acid, curcumin) can also be included.

A further optional component that can be included in the composition isa hydrating agent. Hydrating agents can be defined as agents thatimprove stratum corneum hydration, reduce transepidermal water loss,maintain skin softness and elasticity or have a general soothingfunction in case of dryness and/or irritation. Examples of suchhydrating agents include propylene glycol, glycerol, sorbitol,polyethylene glycol or other agents described in U.S. Pat. No. 5,603,943or US 20070190081. Preferable hydrating agents include, dexpanthenol orpanthothenic acid at a concentration ranging from 0.25-2.5% w/w, morepreferably 0.5-1.5% w/w, more preferably 1% w/w, hyaluronic acid or asalt thereof (such as sodium hyaluronate) at a concentration rangingfrom 0.01-2.5% w/w preferably 0.025-0.5% w/w, more preferably 0.2% w/wor ectoin, a natural compound found in several species of bacteria suchas Ectothiorhodospira halochloris, at a concentration ranging from 0.001to 10% w/w, preferably 0.01-2% w/w, more preferably 0.5-1% w/w.

Respiratory Tract and Respiratory Mucosal-Related Conditions

The methods and compositions of the invention are useful for thetreatment of respiratory tract and/or respiratory mucosal-relatedconditions. Respiratory tract and respiratory mucosal-related conditionsinclude any conditions or disorders with abnormal mucus production,secretion or clearance or inflammation (bacterial, viral, allergic orautoimmune) of the nasal, bronchial and pulmonary mucosa. Examples ofdiseases include acute rhinosinusitis, recurrent acute rhinosinusitis,subacute rhinosinusitis, chronic rhinosinusitis, acute exacerbation ofchronic rhinosinusitis, sinusitis, allergic and non-allergic rhinitis,asthma, cystic fibrosis, chronic obstructive pulmonary disease (COPD),and other diseases of the upper and lower respiratory tract.

The principal functions of the nasal passages are the regulation of theinspiratory air flow, the heating and humidification of inspired air andthe elimination of foreign elements (dust, microbial germs). The ciliateepithelium, which covers the nasal mucosa, plays a vital role in itsdefense from infections. The vibratile cilia of this epithelium areimmersed in protective mucus, which surrounds them, and their purpose isto push this mucous, together with any external impurities, posteriorly,i.e. towards the pharynx, by their motions.

Rhinosinusitis is the inflammation (bacterial, viral, allergic orautoimmune) of the paranasal sinuses. Rhinosinusitis can be classifiedinto several categories: Acute rhinosinusitis—a new infection that maylast up to four weeks and can be subdivided symptomatically into severeand non-severe; Recurrent acute rhinosinusitis—four or more separateepisodes of acute sinusitis that occur within one year; Subacuterhinosinusitis—an infection that lasts between four and 12 weeks, andrepresents a transition between acute and chronic infection; Chronicrhinosinusitis—when the signs and symptoms last for more than 12 weeks;and Acute exacerbation of chronic rhinosinusitis—when the signs andsymptoms of chronic rhinosinusitis exacerbate, but return to baselineafter treatment. Symptoms of rhinosinusitis include nasal congestion,facial pain, headache, fever, and general malaise. Some nonlimitingexamples of inflammatory conditions would include: acute and chronicrhinosinusitis, allergic and non-allergic rhinitis, asthma, cysticfibrosis, chronic obstructive pulmonary disease (COPD), other diseasesof the respiratory tract.

There are several paired paranasal sinuses, including the frontal,ethmoidal, maxillary and sphenoidal sinuses. Rhinosinusitis often occursas part of a spectrum of diseases that affect the respiratory tract andrespiratory mucosal-related conditions are often linked to otherconditions, such as asthma or cystic fibrosis. All forms ofrhinosinusitis may either result in, or be a part of, a generalizedinflammation of the airway, so other airway symptoms, such as cough, maybe associated with it.

Pharmaceutical Compositions

The compositions provided herein are also useful as pharmaceuticalcompositions for treatment of respiratory tract and/or respiratorymucosal-related conditions, such as rhinosinusitis. Pharmaceuticalcompositions can include a pharmaceutically acceptable carrier oradjuvant or can be provided without such a carrier or adjuvant. In oneembodiment, the pharmaceutical composition includes an ionic aqueoussolution, an extract from brown algae, and a pharmaceutically acceptablecarrier or adjuvant. In another embodiment, the pharmaceuticalcomposition includes an ionic aqueous solution, a branched, sulfatedpolysaccharide having an average molecular weight greater than 4 kDa andcomprising L-fucose and sulfate ester groups and a pharmaceuticallyacceptable carrier or adjuvant. The pharmaceutically acceptable carriersor adjuvants useful are conventional. Remington's PharmaceuticalSciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 15thEdition (1975), describes compositions and compositions suitable forpharmaceutical delivery are described herein.

The composition may be contained in any appropriate amount in anysuitable carrier substance, and is generally present in an amount of1-99% by weight of the total weight of the pharmaceutical composition.Pharmaceutical compositions may be formulated according to conventionalpharmaceutical practice (see, e.g., Remington: The Science and Practiceof Pharmacy (20th ed.), ed. A. R. Gennaro, Lippincott Williams &Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J.Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York).

In addition to biologically-neutral carriers, pharmaceuticalcompositions to be administered can contain minor amounts of non-toxicauxiliary substances, such as wetting or emulsifying agents,preservatives, and pH buffering agents and the like, for example sodiumacetate or sorbitan monolaurate.

An exemplary embodiment of a composition for nasal administration caninclude a seawater/saline solution exhibiting an osmolarity valueranging from 250-350 mOsm/L for isotonic, >350 mOsm/L for hypertonic and<250 mOsm/L for hypotonic corresponding to a 10 to about 99.9% weightcontent within the aqueous solution [the corresponding NaCl contents are0.9%, >0.9% and <0.9% NaCl or NaCl equivalents], a branched, sulfatedpolysaccharide having an average molecular weight greater than 4 kDawith L-fucose and sulfate ester groups, and optionally, any or both of ablue/green algae extract and a galactose-based polysaccharide such as acarrageenan.

Another exemplary embodiment of a composition for nasal administrationcan include seawater/saline solution, an extract from brown algae, andblue/green algae extract such as Spirulina, and, optionally, a red algaeextract such as carrageenan.

In certain embodiments, the composition can be packaged in a pressurizedcontainer, equipped with an auto-sterilizing nozzle, with instructionsfor use.

Methods of Delivery

Pharmaceutical compositions described above can be delivered viadifferent methodologies including sprays, irrigation systems (e.g. netipot), syringes or others. The composition may be provided in a dosageform that is suitable for a nasal aerosol or inhalation administrationroute. An exemplary method of administration of the composition caninclude spraying vaporized or nebulized disseminated microparticlesunder an active dynamic pressure.

Suitable aerosol dispensers for use will be apparent to those skilled inthe art, and may vary from simple devices analogous to perfumedispensers to pressurized spray cans and even complex apparatus such asmight be used in hospitals. Whichever device is used it is generallypreferable that it comprises some kind of dosimeter to control theamount of solution administered in one go. One device, which correspondsto a dispenser with a nozzle, effectively incorporates such a dosimeterwithout any specialized adaptation being necessary, the limit stop ofthe depressible spray head fixing the maximum single amount of solutiondispensable at once. Specially developed spray devices may be made witha hand-held device comprising a reservoir of the composition.

Suitable means for dispersing the spray, preferably in aerosol form, areprovided. Examples include pneumatically pressurized devices and devicesemploying pressurized gas forced across the opening of a tube leadinginto the reservoir to create an aerosol, and press-button type deviceswherein the button, when pressed, creates pressure on the surface of theliquid in the reservoir, forcing it up through a tube and through a finenozzle to disperse the solution into an aerosol spray. Other examplesinclude aerosol dispensers, inhalers, pump sprayers, nebulizers (such aspositive pressure nebulizers), and the like. In some embodiments, thedevice used is pre-filled with a composition described herein.

One embodiment would include a multi-dose metered dose spray pumpallowing for spraying of a fixed volume of solution. Alternatively, gasdriven (e.g. nitrogen) devices, such as systems that hold thecomposition separate from the propellant in aluminum or plastic (or anyother type of) bottle. These devices deliver solution at variablediffusion flows and angles when combined with different actuators.Preferred diffusion flows could deliver 0.5-10 ml solution per sprayingsecond at angles of 0-60° C.

The compositions described above can be administered as per physician'sinstructions and depending on the condition. A preferred mode of (nasal)administration would comprise 1-5 sprays per nostril, 1-5 times daily;this could extend to many weeks depending on the condition or symptom tobe treated (e.g. in allergy).

Treatment

The methods described herein provide for treating respiratory tractand/or respiratory mucosa-related conditions or symptoms thereof thatinclude acute and chronic rhinosinusitis, sinusitis, allergic andnon-allergic rhinitis, asthma, cystic fibrosis, chronic obstructivepulmonary disease (COPD), and other diseases of the upper and lowerrespiratory tract. In one embodiment, methods include treating arespiratory tract and/or respiratory mucosa-related condition in asubject in need thereof, by aerosolizing a pharmaceutical composition ina nasal passageway of the subject. Such methods include the step ofadministering to the subject a therapeutic amount a composition hereinsufficient to treat rhinosinusitis, e.g., acute and chronicrhinosinusitis, sinusitis, allergic and non-allergic rhinitis, asthma,cystic fibrosis, chronic obstructive pulmonary disease (COPD), and otherdiseases of the respiratory tract.

The methods and compositions described herein can be administered incombination with one or more agents or separately as successivetreatments. For example, one embodiment describes the compositionadministered in combination as a single treatment with at least one of acorticosteroid, other steroidal compound, anti-histamine, decongestant,bronchodilator, β2-adrenergic agonist, and antimicrobial agent. Inanother embodiment, the composition is administered as an independenttreatment with another, successive treatment including at least one of acorticosteroid, other steroidal compound, anti-histamine, decongestant,bronchodilator, β2-adrenergic agonist, and antimicrobial agent.

It will be appreciated that, although not precluded, treatingrhinosinusitis does not require that rhinosinusitis or symptomsassociated therewith be completely eliminated.

The therapeutic methods described herein (that include prophylactictreatment) in general comprise administration of a therapeuticallyeffective amount of the compositions herein to a subject (e.g., animal,human) in need thereof, including a mammal, particularly a human. Suchtreatment will be suitably administered to subjects, particularlyhumans, suffering from, having, susceptible to, or at risk forrhinosinusitis or inflammatory condition, or symptom thereof.Determination of those subjects “at risk” can be made by any objectiveor subjective determination by a diagnostic test or opinion of a subjector health care provider.

In addition to standard methods of pharmaceutical chemistry andformulations described above, the practice of the compositions andmethods described herein can employ, unless otherwise indicated,conventional techniques of molecular biology (including recombinanttechniques), microbiology, cell biology, biochemistry and immunology,which are well within the purview of the skilled artisan. Suchtechniques are explained fully in the literature, such as, “MolecularCloning: A Laboratory Manual”, second edition (Sambrook, 1989);“Oligonucleotide Synthesis” (Gait, 1984); “Animal Cell Culture”(Freshney, 1987); “Methods in Enzymology” “Handbook of ExperimentalImmunology” (Weir, 1996); “Gene Transfer Vectors for Mammalian Cells”(Miller and Calos, 1987); “Current Protocols in Molecular Biology”(Ausubel, 1987); “PCR: The Polymerase Chain Reaction”, (Mullis, 1994);“Current Protocols in Immunology” (Coligan, 1991). These techniques areapplicable to the production of the polynucleotides and polypeptides ofthe invention, and, as such, may be considered in making and practicingthe invention. Particularly useful techniques for particular embodimentswill be discussed in the sections that follow.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the assay, screening, and therapeutic methods of theinvention, and are not intended to limit the scope of what the inventorsregard as their invention.

Example 1: Formulations of the Invention are Capable of ReducingInflammatory Marker Secretion

FIG. 1 presents data obtained with solutions described in Formulations1-4 in a three-dimensional reconstituted human airway epithelia model(commercially available by Epithelix, Geneva, CH). The airway epitheliaare freshly isolated from nasal biopsies and fully differentiated. Dueto the cells differentiated nature and properties, the model isconsidered a highly physiological system to study effects of appliedagents including cytotoxicity and inflammation. Thus, in the experimentpresented in FIG. 1, fully differentiated human nasal epithelial cellswere reconstituted with a mixture of cells isolated from nasal polyps of14 different normal donors in three-dimensional cultures closelyresembling natural nasal tissues (Huang S et al, The Use of In Vitro 3DCell Models in Drug Development for Respiratory Diseases, InTech. 2011ISBN: 978-953-307-615-7). Each patient cell insert used was washedapically with culture medium 3 days before the experiment to removeaccumulated mucous and to minimize the risk of interference with thetests. To verify that all selected inserts satisfy appropriate qualitycontrols prior to use of test agents, inserts were inspected under themicroscope to verify proper function of the cilia and relevantTrans-Epithelial Electrical Resistance (TEER). Subsequently, LPS wasadded to the culture medium at 0.3 mg/ml in the presence or absence ofdifferent solutions (10 μl applied apicaly two times a day within 8hours) for a total of four days. Culture medium was collected at days 0,1, 2, 3 and 4 and IL-8, a well-established inflammatory marker secretedby airway epithelial cells, was measured by ELISA.

As expected, untreated cultures did not show any significant change ofIL-8 release. LPS greatly stimulated the IL-8 expression in atime-dependent manner, with a peak at Day 3. A hypertonic seawatersolution comprising 2.3% NaCl was capable of reducing the IL-8 releasecaused by LPS. Interestingly, solutions comprising seawater 2.3% NaCland indicated ingredients (UP: Undaria pinnafitida extract, SP:Spirulina platensis extract) (Formulations 1-4) were superior toseawater 2.3% NaCl alone (*p<0.05, **p<0.01, Student's t-test)indicating that algae ingredients potentiate the effect of seawater 2.3%NaCl in a statistically significant manner. It is noted that all testedsolutions were not cytotoxic to the cultured cells (as measured by lackof LDH release in the medium).

Example 2: Formulations of the Invention Reduce Inflammation and ImproveNasal Cell Function in Cultured Epithelial Cells from an Allergic Donor

In the experiments presented in FIG. 2, fully differentiated human nasalepithelial cells were reconstituted with a mixture of inserts derivedfrom an allergic donor (female, 35 years old) in three-dimensionalcultures closely resembling natural nasal tissues (Huang S et al, TheUse of In Vitro 3D Cell Models in Drug Development for RespiratoryDiseases, InTech. 2011 ISBN: 978-953-307-615-7). Cells were washedapically with culture medium 3 days before the experiment to removeaccumulated mucus and to minimize the risk of interference with thetests. Microscopic analysis of cells and relevant Trans-EpithelialElectrical Resistance (TEER) measurements confirmed proper function ofthe cilia prior to the initiation of the experiment.

The experimental design following the aforementioned cell preparationwas as follows: At Day 0, cells were left untreated (negative control)or were treated with 10 μl of Formulation 1, saline control or Cytomix(a mixture of IL-1beta, TNF-alpha and IFN-gamma predicted to exertcytotoxic effects to the nasal epithelium) applied onto the apicalsurface of the epithelium. 24 hours later (Day 1), the effect ofsolutions on Mucociliary Clearance (MCC) was monitored using a Ds-5mccamera (Nikon) connected to a DMIRE2 microscope (Leica). Specifically, 5μM microbeads were added at the apical surface of cell cultures as thesewere incubated with Formulation 1 (or saline control) and 1 minutemovies showing the movement of beads were taken. Subsequent analysesusing the imaging software Image Pro Plus (Mediacy) tracked movements ofbeads and velocity of each particle was calculated in order to determinethe speed of MCC (in μM/sec). Following MCC measurements, 30 μl of 20 mMHCPt (Ammonium hexachloroplatinate; a strong respiratory sensitizer) orsaline was added to cells in the presence of Formulation 1 or salinecontrol. After 16 hours of exposure (Day 2), HCPt solutions were removedand new cell culture medium containing Formulation 1 or saline controlwas added to cells. Treatments with solutions continued twice dailythroughout Day 2 and in Day 3; solutions were removed during at Day 4(replaced with cell culture medium). During this time (Days 1-4),culture media was collected for analysis of inflammatory marker IL-8 byELISA. At Day 4, cilia beating frequency (CBF) measurements (expressedas Hz) were conducted by a dedicated setup including a camera (Sony XCDV60 Firewire), a PCI card and relevant software.

FIG. 2, panel A, shows that Formulation 1 significantly increasedmucociliary clearance in comparison to saline (results represents anaverage value of 200-500 particles tracked). As expected Cytomix had adetrimental effect on the nasal epithelium that resulted in significantreduction of MCC versus baseline. Panel B presents IL-8 secretion dataat Days 1-4. As expected, HCPt efficiently induced inflammation with apeak of IL-8 secretion at Day 4. No IL-8 secretion was observed in cellsreceiving saline instead of HCPt or in untreated cultures. Formulation 1was capable of reducing inflammation induced by respiratory allergenHCPt in these three dimensional cultures from an allergic donor. Panel Cshows CBF measurements at Day 4. As expected cell cultures treated withHCPt had an impaired function as measured by reduced CBF versus control.On the contrary, cultures treated with HCPt that have receivedFormulation 1 had increased CBF values in agreement with protectiveproperties observed in the IL-8 analyses and the improved functionexerted on MCC. Overall, these results suggest that Formulation 1improves nasal function parameters in three dimensional cultures of cellislets generated by an allergic donor and reduces inflammation inducedby a respiratory allergen. It is noted that specific measurements of LDHrelease (in the culture medium) conducted during the course of theseexperiments, proved that Formulation 1 was not cytotoxic to culturedcells (data not shown).

Example 3: Materials and Methods

The following example formulations may be prepared:

Formulation 1

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Purified/distilled/filtered water q.s. 100.0 g

Formulation 2

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Purified/distilled/filtered water q.s. 100.0 g

Formulation 3

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 gPurified/distilled/filtered water q.s. 100.0 g

Formulation 4

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 gPurified/distilled/filtered water q.s. 100.0 g

Formulation 5

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Purified/distilled/filtered waterq.s. 100.0 g

Formulation 6

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Purified/distilled/filtered waterq.s. 100.0 g

Formulation 7

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Dexpanthenol 1.0 g Purified/distilled/filtered water q.s.100.0 g

Formulation 8

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Dexpanthenol 1.0 g Purified/distilled/filtered water q.s.100.0 g

Formulation 9

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Dexpanthenol 1.0 gPurified/distilled/filtered water q.s. 100.0 g

Formulation 10

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Dexpanthenol 1.0 gPurified/distilled/filtered water q.s. 100.0 g

Formulation 11

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Dexpanthenol 1.0 gPurified/distilled/filtered water q.s. 100.0 g

Formulation 12

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Dexpanthenol 1.0 gPurified/distilled/filtered water q.s. 100.0 g

Formulations 1-12 result in hypertonic seawater solutions ofapproximately 2.2-2.5% NaCl.

Formulation 13

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Purified/distilled/filtered water q.s. 100.0 g

Formulation 14

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Purified/distilled/filtered water q.s. 100.0 g

Formulation 15

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Purified/distilled/filtered waterq.s. 100.0 g

Formulation 16

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Dexpanthenol 1.0 g Purified/distilled/filtered water q.s.100.0 g

Formulation 17

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Dexpanthenol 1.0 g Purified/distilled/filtered water q.s.100.0 g

Formulation 18

A seawater solution comprising:

Seawater 28.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Iota-Carrageenan 0.16 g Dexpanthenol 1.0 gPurified/distilled/filtered water q.s. 100.0 g

Formulations 13-18 result in isotonic seawater solutions ofapproximately 0.9% NaCl

Preparation of solutions of Formulations 1-18

Formulations 1-18 are made by charging in the mixer

71.6 or 28.6 g of Seawater  0.1 or 0.3 g of Undaria pinnafitida extract 0.5 g (or not) of Spirulina platensis extract 0.16 g (or not) ofIota-Carrageenan   1 g (or not) of Dexpanthenol

and the remaining purified water and mix for 15 min until a clear,homogeneous solution is obtained.

Formulation 19

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gSeawater (electrodialyzed to 2.2% NaCl) q.s. 100.0 g

Formulation 20

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Seawater (electrodialyzed to 2.2%NaCl) q.s. 100.0 g

Formulation 21

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g Seawater (electrodialyzed to 2.2% NaCl) q.s.100.0 g

Formulation 22

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gSeawater (electrodialyzed to 0.9% NaCl) q.s. 100.0 g

Formulation 23

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g Seawater (electrodialyzed to 0.9% NaCl) q.s.100.0 g

Formulation 24

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gDexpanthenol 1.0 g Seawater (electrodialyzed to 2.2% NaCl) q.s. 100.0 g

Formulation 25

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Dexpanthenol 1.0 g Seawater(electrodialyzed to 2.2% NaCl) q.s. 100.0 g

Formulation 26

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g Dexpanthenol 1.0 g Seawater (electrodialyzed to2.2% NaCl) q.s. 100.0 g

Formulation 27

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gDexpanthenol 1.0 g Seawater (electrodialyzed to 0.9% NaCl) q.s. 100.0 g

Formulation 28

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g Dexpanthenol 1.0 g Seawater (electrodialyzed to0.9% NaCl) q.s. 100.0 g

Preparation of solutions of Formulations 19-28

Following the method of U.S. Pat. No. 6,451,352, these formulations maybe prepared by electrodialyzing seawater to the desired osmotic content,then adding and mixing the additional ingredients. An alternative methodis to mixing all the ingredients together and then electrodialyze.

Formulation 29

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Spirulina platensisextract 0.5 g Eucalyptus oil 0.0075 g Spearmint oil 0.015 g Thymeextract 0.1 g 1,3 propanediol 5 g Glycerin 3 grPurified/distilled/filtered water q.s. 100.0 g

Formulation 30

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Spirulina platensisextract 0.5 g Eucalyptus oil 0.0075 g Spearmint oil 0.015 g Thymeextract 0.1 g 1,3 propanediol 5 g Glycerin 3 grPurified/distilled/filtered water q.s. 100.0 g

Formulation 31

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.1 g Eucalyptus oil 0.0075g Spearmint oil 0.015 g Thyme extract 0.1 g 1,3 propanediol 5 g Glycerin3 gr Purified/distilled/filtered water q.s. 100.0 g

Formulation 32

A seawater solution comprising:

Seawater 71.6 g Undaria pinnafitida extract 0.3 g Eucalyptus oil 0.0075g Spearmint oil 0.015 g Thyme extract 0.1 g 1,3 propanediol 5 g Glycerin3 gr Purified/distilled/filtered water q.s. 100.0 g

Preparation of solutions of Formulations 29-32

Formulations 29-32 are made by charging in the mixer the followingingredients to form a first mix:

71.6 or 28.6 g of Seawater  0.1 or 0.3 g of Undaria pinnafitida extract 0.5 g (or not) of Spirulina platensis extract 0.16 g (or not) ofIota-Carrageenan  0.1 g (or not) of Thyme extract

A second mix containing (in a small volume of purified water):

0.0075 g of Eucalyptus oil  0.015 g of Spearmint oil    5 gr of 1,3propanediol

Then addition of glycerin in the 2^(nd) mix:

3 gr of Glycerin

The second mix is then added to the first mix under continuous andintense stirring over 30 minutes and purified water is added to a finalweight of 100 grams.

Formulation 33

A seawater solution comprising:

Undaria pinnafitida extract 0.1 g Spirulina platensis extract 0.5 gEucalyptus oil 0.0075 g Spearmint oil 0.015 g Thyme extract 0.1 g 1,3propanediol 5 g Glycerin 3 gr Seawater (electrodialyzed to 2.2% NaCl)q.s. 100.0 g

Formulation 34

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gEucalyptus oil 0.0075 g Spearmint oil 0.015 g Thyme extract 0.1 g 1,3propanediol 5 g Glycerin 3 gr Seawater (electrodialyzed to 2.2% NaCl)q.s. 100.0 g

Formulation 35

A seawater solution comprising:

Undaria pinnafitida extract 0.1 g Eucalyptus oil 0.0075 g Spearmint oil0.015 g Thyme extract 0.1 g 1,3 propanediol 5 g Glycerin 3 gr Seawater(electrodialyzed to 2.2% NaCl) q.s. 100.0 g

Formulation 36

A seawater solution comprising:

Undaria pinnafitida extract 0.3 g Eucalyptus oil 0.0075 g Spearmint oil0.015 g Thyme extract 0.1 g 1,3 propanediol 5 g Glycerin 3 gr Seawater(electrodialyzed to 2.2% NaCl) q.s. 100.0 g

Preparation of solutions of Formulations 33-36

Following the method of U.S. Pat. No. 6,451,352, these formulations maybe prepared by electrodialyzing seawater to the desired osmolarity.Then, a first mix is made by adding algae ingredients and Thyme extractin electodialyzed water. A second mix in a smaller volume ofelectrodialyzed seawater is created and mixed, glycerin is then addedwith intense stirring over 30 minutes, then all mixes are pooled to thefinal volume adjusting the final osmolarity with electrodialyzedseawater of the appropriate osmolarity.

Seawater Used to Prepare Samples

The seawater solution to be used for the preparation of these solutionsis preferably derived from dedicated seawater collection sites from adepth of 5-10 meters in zones characterized by the presence of strongcurrents (which guarantee continuous replenishment of fresh water). Theseawater, upon collection, may be typically filtered to remove organiccontaminants and/or microorganisms.

A collection site is at the Cancale Bay, Bretagne, France, see EP0909184. Specifications of the seawater collected at Cancale Bay arepresented in Table 4.

TABLE 4 Specifications of Seawater. CONTROL SPECIFICATIONS METHODAppearance Colorless clear liquid Organoleptic Odor CharacteristicOrganoleptic pH (20° C.) 7.5-8.5 Eur. Pharm. 7/2.2.3 Density (20° C.)0.975-1.078 g/ml Eur. Pharm. 7/2.2.5 Arsenic <2 ppm Eur. Pharm. 7/2.4.2Heavy metals <1 ppm Eur. Pharm. 7/2.4.8 Osmolality 1000-1200 mOsm/L Eur.Pharm. 7/2.2.35 TOC <50 ppm Eur. Pharm. 7/2.2.44 NαCl Content 29.0-34.0g/l Volhard Method Total hydrocarbon <1 mg/l St. Methods/503A Totalaerobic microbial <1000 cfu/ml Eur. Pharm. 7/2.6.12 Count Yeasts andmoulds Count <100 cfu/ml Eur. Pharm. 7/2.6.12 Enterobacteria <10 cfu/mlEur. Pharm. 7/2.6.13 Staphylococcus aureus Absence/10 ml Eur. Pharm.7/2.6.13 Thermotolerant coliforms Absence/10 ml Eur. Pharm. 7/2.6.13Pseudomonas Absence/10 ml Eur. Pharm. 7/2.6.13 aeruginosa

Formulation 37

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g 1% saline solution q.s. 100 g

Formulation 38

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 g 1%saline solution q.s. 100 g

Formulation 39

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g 1% saline solution q.s. 100 g

Formulation 40

A saline solution comprising

Sodium Chloride 3 g Undaria pinnafitida extract 0.3 g Spirulinaplatensis extract 0.5 g Purified water q.s. 100 g

Formulation 41

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g Dexpanthenol 1.0 g 1% salinesolution q.s. 100 g

Formulation 42

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gDexpanthenol 1.0 g 1% saline solution q.s. 100 g

Formulation 43

A saline solution comprising

Magnesium Chloride 2 g Magnesium Bromide 0.05 g Magnesium sulfate 0.05 gPotassium Chloride 1.00 g Calcium Chloride 0.05 g Sodium Carbonate 0.05g Undaria pinnafitida extract 0.3 g Spirulina platensis extract 0.5 gIota-Carrageenan 0.16 g Dexpanthenol 1.0 g 1% saline solution q.s. 100 g

Formulation 44

A saline solution comprising

Sodium Chloride 3 g Undaria pinnafitida extract 0.3 g Spirulinaplatensis extract 0.5 g Dexpanthenol 1.0 g Purified water q.s. 100 g

Saline Samples (Formulations 37-44)

The preparation of saline samples is according to U.S. Pat. No.7,541,052 (mixing of all ingredients under stirring till the solutionbecomes clear and homogenous).

After preparation of any of the formulations, the solutions may befiltered using appropriate filters (e.g. 0.22 μM), sterilized bystandard methodologies (e.g. γ-irradiation) and used in the preparationsof devices delivering the solutions.

OTHER EMBODIMENTS

From the foregoing description, it will be apparent that variations andmodifications may be made to the invention described herein to adopt itto various usages and conditions. Such embodiments are also within thescope of the following claims.

The recitation of a listing of elements in any definition of a variableherein includes definitions of that variable as any single element orcombination (or subcombination) of listed elements. The recitation of anembodiment herein includes that embodiment as any single embodiment orin combination with any other embodiments or portions thereof.

All patents and publications mentioned in this specification are hereinincorporated by reference to the same extent as if each independentpatent and publication was specifically and individually indicated to beincorporated by reference.

1. A method for treating a respiratory tract and/or respiratorymucosal-related condition in a subject in need thereof, comprisingaerosolizing a pharmaceutical composition in a nasal passageway of thesubject wherein the pharmaceutical composition comprises: an ionicaqueous solution; and a sulfated polysaccharide having an averagemolecular weight greater than 4 kDa and comprising L-fucose and sulfateester groups.
 2. The method of claim 1, wherein the subject is a humansubject.
 3. The method of claim 1, wherein the respiratory tract and/orrespiratory mucosal-related condition is selected from the groupconsisting of acute and chronic rhinosinusitis, sinusitis, allergic andnon-allergic rhinitis, asthma, cystic fibrosis, chronic obstructivepulmonary disease (COPD), and other diseases of the respiratory tract.4. The method of claim 3, wherein the respiratory tract and/orrespiratory mucosal-related condition is rhinosinusitis.
 5. The methodof claim 3, wherein the respiratory tract and/or respiratorymucosal-related condition is allergic rhinitis.
 6. The method of claim1, wherein the sulfated polysaccharide is derived from an extractisolated from brown algae.
 7. The method of claim 6, wherein the brownalgae is fucoidan.
 8. The method of claim 6, wherein the brown algae isUndaria pinnatifida.
 9. The method of claim 1, wherein thepharmaceutical composition further comprises an extract of blue/greenalgae.
 10. The method of claim 1, wherein the pharmaceutical compositionfurther comprises at least one of the following: a sulfated,galactose-based polysaccharide having an average molecular weightgreater than 15 kDa; at least one substance selected from exogenoussalts, anti-microbial agents, amino acids, and essential oils; thymeextract; and at least one hydrating agent.
 11. The method of claim 1,wherein the pharmaceutical composition further comprises at least onesubstance selected from corticosteroids, other steroidal compounds,anti-histamines, decongestants, bronchodilators, β2-adrenergic agonists,and antibiotics.
 12. The method of claim 1, wherein the sulfatedpolysaccharide is in a range of 0.1 to 10% weight content of thepharmaceutical composition and when present in the pharmaceuticalcomposition, the extract of a blue/green algae is in the range of 0.1 to10% weight content of the pharmaceutical composition and thegalactose-based polysaccharide is in the range of 0.1 to 10% weightcontent of the pharmaceutical composition.
 13. The method of claim 9,wherein the extract of blue/green algae is isolated from Spirulinaplatensis.
 14. The method of claim 10, wherein the galactose-basedpolysaccharide is a carrageenan or wherein the galactose-basedpolysaccharide is derived from an extract isolated from red algae. 15.The method of claim 14, wherein the carrageenan is selected from thegroup consisting of iota-carrageenan, kappa-carrageenan, andlambda-carrageenan (e.g. iota-carrageenan).
 16. The method of claim 14,wherein the red algae is selected from the group consisting of Gigartinastellate, Chondrus chrispus, Kappaphycus cottonii.
 17. The method ofclaim 10, wherein the hydrating agent is dexpanthenol or panthothenicacid and wherein dexpanthenol or panthothenic acid is present at aconcentration ranging from 0.25-2.5% w/w.
 18. The method of claim 1,wherein ionic aqueous solution is in a range of 10 to 99.9% weightcontent of the composition.
 19. The method of claim 1, wherein the ionicaqueous solution comprises seawater.
 20. The method of claim 1, whereinthe ionic aqueous solution comprises saline.